Impact of Glucocorticoid on a Cellular Model of Parkinson’s Disease: Oxidative Stress and Mitochondrial Function

Stress seems to contribute to the neuropathology of Parkinson’s disease (PD), possibly by dysregulation of the hypothalamic–pituitary–adrenal axis. Oxidative distress and mitochondrial dysfunction are key factors involved in the pathophysiology of PD and neuronal glucocorticoid-induced toxicity. Animal PD models have been generated to study the effects of hormonal stress, but no in vitro model has yet been developed. Our aim was to examine the impact of corticosterone (CORT) administration on a dopaminergic neuronal cell model of PD induced by the neurotoxin MPP+, as a new combined PD model based on the marker of endocrine response to stress, CORT, and oxidative-mitochondrial damage. We determined the impact of CORT, MPP+ and their co-incubation on reactive oxygen species production (O2−•), oxidative stress cellular markers (advanced-oxidation protein products and total antioxidant status), mitochondrial function (mitochondrial membrane potential and mitochondrial oxygen consumption rate) and neurodegeneration (Fluoro-Jade staining). Accordingly, the administration of MPP+ or CORT individually led to cell damage compared to controls (p < 0.05), as determined by several methods, whereas their co-incubation produced strong cell damage (p < 0.05). The combined model described here could be appropriate for investigating neuropathological hallmarks and for evaluating potential new therapeutic tools for PD patients suffering mild to moderate emotional stress.

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Regulation of Actg1 and Gsta2 is possible mechanism by which capsaicin alleviates apoptosis in cell model of 6-OHDA-induced Parkinson's disease

Bioscience Reports ◽

10.1042/bsr20191796 ◽

2020 ◽

Vol 40 (6) ◽

Cited By ~ 1

Author(s):

Jiahui Liu ◽

Hong Liu ◽

Zhenxiang Zhao ◽

Jianfeng Wang ◽

Dandan Guo ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Cell Model ◽

Cell Damage ◽

Glutathione S Transferase ◽

Expression Arrays ◽

Transcriptional Changes ◽

Molecular Mechanism Of Action ◽

Disease Cell

Abstract The present study aimed to identify the gene expression changes conferred by capsaicin in the cell model of 6-OHDA-induced Parkinson's disease, to disclose the molecular mechanism of action of capsaicin. We used capsaicin-treated and paraffin-embedded wax blocks containing substantia nigra tissue from 6-OHDA-induced Parkinson's disease rats to analyze transcriptional changes using Affymetrix GeneChip Whole Transcript Expression Arrays. A total of 108 genes were differentially expressed in response to capsaicin treatment, and seven of these genes were selected for further analysis: Olr724, COX1, Gsta2, Rab5a, Potef, Actg1, and Acadsb, of which Actg1 (actin gamma 1) was down-regulated and Gsta2 (Glutathione S-transferase alpha 2) was up-regulated. We successfully overexpressed Actg1 and Gsta2 in vitro. CCK-8 detection and flow cytometry demonstrated that overexpression of Actg1 and Gsta2 increased apoptosis in the 6-OHDA-induced Parkinson's disease cell model. The imbalance between Actg1 and Gsta2 may be one of the mechanisms of cell damage in Parkinson's disease (PD). Capsaicin can protect the cells and reduce the apoptosis rate by regulating Actg1 and Gsta2.

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TSH Combined with TSHR Aggravates Diabetic Peripheral Neuropathy by Promoting Oxidative Stress and Apoptosis in Schwann Cells

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/2482453 ◽

2021 ◽

Vol 2021 ◽

pp. 1-23

Author(s):

Jingwen Fan ◽

Qi Pan ◽

Qun Gao ◽

Wenqing Li ◽

Fei Xiao ◽

...

Keyword(s):

Oxidative Stress ◽

Peripheral Neuropathy ◽

Diabetic Peripheral Neuropathy ◽

Cell Model ◽

Cell Damage ◽

Motor Nerve ◽

Thyroid Stimulating Hormone ◽

Motor Nerve Conduction Velocity ◽

The Impact

Subclinical hypothyroidism (SCH) is associated with diabetic peripheral neuropathy (DPN); however, the mechanism underlying this association remains unknown. This study is aimed at examining neurofunctional and histopathological alterations in a type 2 diabetes (T2DM) mouse model of SCH and investigating the impact of thyroid-stimulating hormone (TSH) in an in vitro DPN cell model established using RSC96 cells under high glucose (HG) and palmitic acid (PA) stimulation. Our results indicated that T2DM, in combination with SCH, aggravated abnormal glucose and lipid metabolism in T2DM and dramatically destroyed the peripheral nervous system by increasing paw withdrawal latency, decreasing motor nerve conduction velocity, and exacerbating ultrastructural deterioration of the damaged sciatic nerve caused by diabetes. Furthermore, the results of our in vitro experiments showed that TSH intensified HG/PA-induced RSC96 cell damage by inducing oxidative stress, mitochondrial dysfunction, and apoptosis. More importantly, TSHR knockout or inhibition of PA-induced TSHR palmitoylation could alleviate the apoptosis induced by TSH. Overall, in this study, the novel mechanisms by which TSH, as an independent risk factor for DPN progression, aggravating Schwann cell apoptosis and demyelination, are elucidated. These findings indicate that TSHR could be a potential target for both the prevention and treatment of DPN and, possibly, other microvascular diseases, and have implication in the clinical management of patients with DPN.

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LncRNA SOX2-OT Participates in Parkinson’s Disease Through Regulating miRNA-942-5p/NAIF1 Axis

10.21203/rs.3.rs-763972/v1 ◽

2021 ◽

Author(s):

Yabi Guo ◽

Yanyang Liu ◽

Hong Wang ◽

Peijun Liu

Keyword(s):

Oxidative Stress ◽

Parkinson’S Disease ◽

Parkinson's Disease ◽

Neuronal Apoptosis ◽

Cell Model ◽

Luciferase Reporter ◽

Sod Activity ◽

Software Analysis ◽

Direct Target

Abstract Parkinson’s disease (PD) is a neurodegenerative disease. Studies have shown that lncRNA SOX2-OT was highly expressed in PD patients, but its specific functions and mechanisms still need further research. This study aimed to explore whether lncRNA SOX2-OT could regulate oxidative stress, inflammation and neuronal apoptosis in PD in vitro model and explored the underlying mechanism. An in vitro PD cell model was induced by 1-methyl-4-phenylpyridinium (MPP+). The results of the biological software analysis and luciferase reporter assay indicated that miR-942-5p was a direct target of lncRNA SOX2-OT, and NAIF1 was a direct target of miR-942-5p. Experiments showed that the expression levels of lncRNA SOX2-OT and NAIF1 were increased, and miR-942-5p expression was decreased in SH-SY5Y cells following MPP+ treatment. In addition, MPP+ treatment reduced SH-SY5Y cell viability, induced apoptosis, increased cleaved-Caspase3 protein expression, and increased cleaved-Caspase3/Caspase3 ratio, increased LDH viability, and increased the levels of TNF-α, IL-1β and ROS in SH-SY5Y cells, reduced SOD activity, however, all these effects were inhibited by SOX2-OT-siRNA, and these inhibitions were reversed by miR-942-5p inhibitor. Moreover, the protective role of miR-942-5p mimic in MPP+ induced SH-SY5Y cells was significantly eliminated by NAIF1-plasmid. In summary, this study confirmed that lncRNA SOX2-OT regulated oxidative stress, inflammation and neuronal apoptosis via directly regulating the miR-942-5p/NAIF1 signal axis, and then participated in the occurrence and development of PD. These data provide a new potential targets for PD diagnosis and treatment.

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Mathematical modeling of the role of α-synuclein and dopamine in cell death in Parkinson’s disease provides the molecular basis to the toxicity of different point mutations

10.1101/714600 ◽

2019 ◽

Author(s):

Gourvendu Saxena ◽

Utkarsh Khandelwal ◽

Mukesh Doble

Keyword(s):

Oxidative Stress ◽

Experimental Data ◽

Parkinson’S Disease ◽

Parkinson's Disease ◽

Endoplasmic Reticulum ◽

Cell Death ◽

Cell Survival ◽

Neuronal Cell ◽

Human Neuroblastoma

ABSTRACTDifferent types of α-synuclein and non-β-amyloid component (NAC) peptides have been shown to induce cell death, with varying degree of toxicity, in various in vitro experiments. Oxidative stress has also been associated and proved to be involved in the pathogenesis of neuronal cell death in Parkinson’s disease. Oxidative stress has been shown to accelerate the aggregation of α-synuclein in vitro and in resent studies α-synuclein has been shown to increase oxidative stress. Thus it seems like a vicious cycle, one promoting the other.In this present work we have modeled the α-synuclein pathway to increase cytoplasmic Dopamine concentration, and thereby increasing the Reactive Oxygen Species (ROS) level of the cell, which consequently results in cell death. This model relates the α-synuclein concentration with the fractional cell survival and provides insight of crucial reaction(s) of α-synuclein which promote cell death. It predicts the toxicity of the type of α-synuclein and also explains the pattern of cell death with increasing concentration of α-synuclein. First we modeled a part of the pathway i.e. from dopamine to cell death. The results were compared with experimental data available for PC12 neuronal cell line. Then modeling of full pathway was done and the results were compared with experimental data available for Human neuroblastoma SH-SY5Y cells. It is predicted from this model that higher the auto catalysis of dopamine, higher is the cell death. Interestingly, the model predicts that NAC (1-18) not only hinders the vesicles coming from Endoplasmic Reticulum, to fuse with Golgi bodies, but also reduces the synthesis of Dopamine and the formation of vesicles from Endoplasmic Reticulum. The model is generalized and can predict the toxicity of any protein which impedes the early secretary pathway in dopaminergic cells and also the cell survival pattern with increasing concentration of the protein.

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ADP/ATP Translocase 1 Protects Against an A-Synuclein-Associated Neuronal Cell Damage in Parkinson's Disease Model

10.21203/rs.3.rs-528268/v1 ◽

2021 ◽

Author(s):

Wenyong Ding ◽

Minghua Qi ◽

Li Ma ◽

Xuefei Xu ◽

Yingfei Chen ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Western Blot ◽

Drug Targets ◽

Neurodegenerative Disorder ◽

Cell Model ◽

Cell Damage ◽

Neuronal Cell ◽

Protein Aggregates ◽

Study Results

Abstract Background: ADP/ATP translocase 1 (ANT1) is involved in the exchange of cytosolic ADP and mitochondrial ATP, and its defection plays an important role in mitochondrial pathogenesis. To reveal an etiological implication of ANT1 for Parkinson’s disease (PD), a neurodegenerative disorder, a mouse model treated with 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine and neuroblasma cell model induced by 1-methyl-4-pehny1-pyridine were utilized in this study. Results: The tissue-specific abundance in ANT1 in mouse brains was accessed using the analysis of Western blot and immunohistochemistry. Down-regulated soluble ANT1 was found to be correlated with PD, and ANT1 was associated with PD pathogenesis via forming protein aggregates with α-synuclein. This finding was confirmed at cellular level using neuroblasma cell models. Protein interaction assay, coupled with the analysis of LC-MS/MS, silver-stained SDS-PAGE and Western blot against anti-ANT1 antibody respectively, illustrated the interaction of ANT1 with α-synuclein using the expressed α-synuclein as a bite. Additionally, a significant increasing ROSs was detected in PD-like cells. Conclusions: his study indicated that ANT1 was a potentially causative factor of PD, and led to neuropathogenic injury via promoting the formation of protein aggregates with α-synuclein. This investigation potentially promotes an innovative understanding of ANT1 on the etiology of PD and provides valuable information on developing potential drug targets in PD treatment or reliable biomarkers in PD prognostication.

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MiR-30a-5p regulates GLT-1 function via a PKCα-mediated ubiquitin degradation pathway in a mouse model of Parkinson’s disease

10.21203/rs.2.24022/v1 ◽

2020 ◽

Author(s):

Xingjun Meng ◽

Jianping Zhong ◽

Chong Zeng ◽

Ken Kin Lam Yung ◽

Xiuping Zhang ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Mouse Model ◽

Cell Model ◽

Glutamate Transporters ◽

Acid Uptake ◽

Dependent Manner ◽

The Impact

Abstract Background:Glutamate excitotoxicity caused by dysfunctional glutamate transporters plays an important role in the pathogenesis of Parkinson’s disease (PD); however, the mechanisms that underlie the regulation of glutamate transporters in PD are still not fully elucidated. MicroRNAs have been reported to play key roles in regulating the translation of glutamate-transporter mRNA. Methods: We established model of PD 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice in vivo and 1-methyl-4-phenylpyridinium (MPP+) treated astrocyte in vitro. Stereotaxic injection of shRNA in mouse, and miRNA inhibitor/mimic, or antagonist/agonist treated the cell model, Behavioral experiments, glutamic acid uptake, transport activity of synaptosomes, underlying mechanisms and the impact on neuronal survival were assessed.Results We demonstrated that short-hairpin RNA-mediated knockdown of miR-30a-5p ameliorated motor deficits and pathological changes like astrogliosis and reactive microgliosis in a mouse model of PD. Western blotting and immunofluorescent labeling revealed that miR-30a-5p suppressed the expression and function of GLT-1 in MPTP-treated mice and specifically in astrocytes treated with (cell model of PD). Conclusion Both in vitro and in vivo, we found that miR-30a-5p knockdown promoted glutamate uptake and increased GLT-1 expression by hindering GLT-1 ubiquitination and subsequent degradation in a PKCα-dependent manner. Therefore, miR-30a-5p represents a potential therapeutic target for the treatment of PD.

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Caspase-1 causes truncation and aggregation of the Parkinson’s disease-associated protein α-synuclein

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1610099113 ◽

2016 ◽

Vol 113 (34) ◽

pp. 9587-9592 ◽

Cited By ~ 92

Author(s):

Wei Wang ◽

Linh T. T. Nguyen ◽

Christopher Burlak ◽

Fariba Chegini ◽

Feng Guo ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Cell Model ◽

Lewy Bodies ◽

Neuronal Cell ◽

Specific Chemical ◽

Caspase 1 ◽

In Vitro Characterization ◽

Induced Aggregation

The aggregation of α-synuclein (aSyn) leading to the formation of Lewy bodies is the defining pathological hallmark of Parkinson’s disease (PD). Rare familial PD-associated mutations in aSyn render it aggregation-prone; however, PD patients carrying wild type (WT) aSyn also have aggregated aSyn in Lewy bodies. The mechanisms by which WT aSyn aggregates are unclear. Here, we report that inflammation can play a role in causing the aggregation of WT aSyn. We show that activation of the inflammasome with known stimuli results in the aggregation of aSyn in a neuronal cell model of PD. The insoluble aggregates are enriched with truncated aSyn as found in Lewy bodies of the PD brain. Inhibition of the inflammasome enzyme caspase-1 by chemical inhibition or genetic knockdown with shRNA abated aSyn truncation. In vitro characterization confirmed that caspase-1 directly cleaves aSyn, generating a highly aggregation-prone species. The truncation-induced aggregation of aSyn is toxic to neuronal culture, and inhibition of caspase-1 by shRNA or a specific chemical inhibitor improved the survival of a neuronal PD cell model. This study provides a molecular link for the role of inflammation in aSyn aggregation, and perhaps in the pathogenesis of sporadic PD as well.

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Neuroprotective Effect of Coptis chinensis in MPP+ and MPTP-Induced Parkinson’s Disease Models

The American Journal of Chinese Medicine ◽

10.1142/s0192415x16500506 ◽

2016 ◽

Vol 44 (05) ◽

pp. 907-925 ◽

Cited By ~ 15

Author(s):

Thomas Friedemann ◽

Yue Ying ◽

Weigang Wang ◽

Edgar R. Kramer ◽

Udo Schumacher ◽

...

Keyword(s):

Oxidative Stress ◽

Parkinson’S Disease ◽

Parkinson's Disease ◽

Neurodegenerative Diseases ◽

Cell Model ◽

Neuroprotective Effect ◽

Treatment Options ◽

Coptis Chinensis

The rhizome of Coptis chinensis is commonly used in traditional Chinese medicine alone or in combination with other herbs to treat diseases characterized by causing oxidative stress including inflammatory diseases, diabetes mellitus and neurodegenerative diseases. In particular, there is emerging evidence that Coptis chinensis is effective in the treatment of neurodegenerative diseases associated with oxidative stress. Hence, the aim of this study was to investigate the neuroprotective effect of Coptis chinensis in vitro and in vivo using MPP[Formula: see text] and MPTP models of Parkinson’s disease. MPP[Formula: see text] treated human SH-SY5Y neuroblastoma cells were used as a cell model of Parkinson’s disease. A 24[Formula: see text]h pre-treatment of the cells with the watery extract of Coptis chinensis significantly increased cell viability, as well as the intracellular ATP concentration and attenuated apoptosis compared to the MPP[Formula: see text] control. Further experiments with the main alkaloids of Coptidis chinensis, berberine, coptisine, jaterorrhizine and palmatine revealed that berberine and coptisine were the main active compounds responsible for the observed neuroprotective effect. However, the full extract of Coptis chinensis was more effective than the tested single alkaloids. In the MPTP-induced animal model of Parkinson’s disease, Coptis chinensis dose-dependently improved motor functions and increased tyrosine hydroxylase-positive neurons in the substantia nigra compared to the MPTP control. Based on the results of this work, Coptis chinensis and its main alkaloids could be considered potential candidates for the development of new treatment options for Parkinson’s disease.

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ADP/ATP translocase 1 protects against an α-synuclein-associated neuronal cell damage in Parkinson’s disease model

Cell & Bioscience ◽

10.1186/s13578-021-00645-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Wenyong Ding ◽

Minghua Qi ◽

Li Ma ◽

Xuefei Xu ◽

Yingfei Chen ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Western Blot ◽

Drug Targets ◽

Neurodegenerative Disorder ◽

Cell Model ◽

Cell Damage ◽

Neuroblastoma Cell ◽

Neuronal Cell ◽

Protein Aggregates

Abstract Background ADP/ATP translocase 1 (ANT1) is involved in the exchange of cytosolic ADP and mitochondrial ATP, and its defection plays an important role in mitochondrial pathogenesis. To reveal an etiological implication of ANT1 for Parkinson’s disease (PD), a neurodegenerative disorder, a mouse model treated with 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine and neuroblastoma cell model induced by 1-methyl-4-pehny1-pyridine were utilized in this study. Results The tissue-specific abundance in ANT1 in mouse brains was accessed using the analysis of Western blot and immunohistochemistry. Down-regulated soluble ANT1 was found to be correlated with PD, and ANT1 was associated with PD pathogenesis via forming protein aggregates with α-synuclein. This finding was confirmed at cellular level using neuroblastoma cell models. ANT1 supplement in neuronal cells revealed the protective roles of ANT1 against cytotoxicity caused by MPP+. Protein interaction assay, coupled with the analysis of LC-MS/MS, silver-stained SDS-PAGE and Western blot against anti-ANT1 antibody respectively, illustrated the interaction of ANT1 with α-synuclein using the expressed α-synuclein as a bite. Additionally, a significant increasing ROSs was detected in the MPP+-treated cells. Conclusions This study indicated that ANT1 was a potentially causative factor of PD, and led to neuropathogenic injury via promoting the formation of protein aggregates with α-synuclein. This investigation potentially promotes an innovative understanding of ANT1 on the etiology of PD and provides valuable information on developing potential drug targets in PD treatment or reliable biomarkers in PD prognostication.

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A novel nanoparticle system targeting damaged mitochondria for the treatment of Parkinson's disease

10.21203/rs.3.rs-989619/v1 ◽

2021 ◽

Author(s):

Yue Chen ◽

Lina Yu ◽

Jinyu Zhang ◽

Lifen Yao ◽

Hongji Yan ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Mitochondrial Function ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Mitochondrial Damage ◽

Irreversible Damage ◽

Novel Strategy

Abstract BackgroundMitochondrial damage is one of the primary causes of neuronal cell death in Parkinson's disease (PD). In PD patients, the mitochondrial damage can be repaired or irreversible. Therefore, mitochondrial damage repair becomes a promising strategy for PD treatment.MethodsWe use hyaluronic acid nanoparticles (HA-NPs) of different molecular weights to protect the mitochondria and salvages the mild and limited damage in mitochondria. Our HA-NPs with 2,190 kDa HA can improve the mitochondrial function of SH-SY5Y cells and PTEN induced putative kinase 1 (PINK1) knockout mouse embryo fibroblast (MEF) cells. In cases of irreversible damage, we use NPs with ubiquitin specific peptidase 30 (USP30) siRNA to promote mitophagy. Meanwhile, by adding PINK1 antibodies, our NPs can selectively target the irreversibly damaged mitochondria, preventing the excessive clearance of healthy mitochondria.ResultsOur HA-NPs with 2,190 kDa HA can protect the mitochondria and salvage the mitochondrial function of the mild and limited damage conditions in both SH-SY5Y cells and PINK1 knockout MEF cells. NPs with USP30 siRNA and PINK1 antibodies can selectively target and promote the clearance of irreversibly damaged mitochondria both in vitro and in vivo.ConclusionsWe successfully designed and developed NPs for the treatment of PD that can target different stages of mitochondrial damage. This strategy is expected to overcome the problems associated with the excessive clearance of healthy mitochondria and has great potential for clinical applications. Our study provides a novel strategy for PD treatment.

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